Transcriptional induction of the human renin gene by cyclic AMP requires cyclic AMP response element-binding protein (CREB) and a factor binding a pituitary …

S GERMAIN, T KONOSHITA, J PHILIPPE… - Biochemical …, 1996 - portlandpress.com
S GERMAIN, T KONOSHITA, J PHILIPPE, P CORVOL, F PINET
Biochemical Journal, 1996portlandpress.com
To delineate the cis-acting elements of the proximal promoter responsible for cyclic AMP
(cAMP)-induced human renin gene transcription, 5´-flanking regions of the human renin
gene were fused to a luciferase reporter gene and transfected in chorionic cells. Forskolin
treatment induced the expression of luciferase by 2.4-fold when the reporter plasmid
contained the promoter region (-582 to+ 16). Mutation or deletion of the cAMP response
element (CRE) diminished (1.7-fold) but did not abolish cAMP-induced transcription …
To delineate the cis-acting elements of the proximal promoter responsible for cyclic AMP (cAMP)-induced human renin gene transcription, 5´-flanking regions of the human renin gene were fused to a luciferase reporter gene and transfected in chorionic cells. Forskolin treatment induced the expression of luciferase by 2.4-fold when the reporter plasmid contained the promoter region (-582 to +16). Mutation or deletion of the cAMP response element (CRE) diminished (1.7-fold) but did not abolish cAMP-induced transcription, demonstrating that the (-582 to -145) region containing the CRE and the region (-145 to -38) containing a Pit-1 (pituitary-specific trans-acting factor) site were both necessary for cAMP maximal induction. To study the molecular events mediating the cAMP induction, DNase I footprinting and electromobility shift assays (EMSAs) were performed with renin-producing chorionic cell and kidney cortex cell nuclear extracts, showing that the CRE-binding protein (CREB) interacts with the CRE and that tissue-specific factors, distinct from Pit-1, specifically bind the renin Pit-1 motif. Taken together, these results demonstrate that the cAMP response of the human renin gene may involve CREB binding the CRE and tissue-specific factors, different from Pit-1, that interact with the Pit-1 response DNA elements.
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